Methods and device for adhesive control of internal cell organisation

Abstract

The present invention relates to methods and devices for adhering cells in a specific and predetermined position with an adhesive control of internal cell organisation, methods for preparing such devices, methods for studying modifications of cell shape and global internal cell organization such as the distribution of cellular compartments, centrosome centering, spindle orientation, internal compartimentalization and internal transports, methods for screening compounds of interest which enhance or inhibit specific cell functions.

Description

FIELD OF THE INVENTION [0001] The present invention relates to methods and devices for adhering cells in a specific and predetermined position with an adhesive control of internal cell organistation, methods for preparing such devices, methods for studying modifications of cell shape and global internal cell organization such as the distribution of cellular compartments, centrosome centering, spindle orientation, internal compartimentalization and internal transports, methods for screening compounds of interest which enhance or inhibit specific cell functions. BACKGROUND OF THE INVENTION [0002] High throughput cell-based phenotypic screening becomes necessary to take advantage of the wealth of data obtained from systematic genome sequencing. Genome wide gene silencing by siRNA is now possible on cultured cells. Alternatively, one would like to rapidly identify biologically active compounds from drugs libraries able to enhance or inhibit specific cell functions. The aim is thus to ca...

AI Technical Summary

Benefits of technology

[0010] In the present invention, the inventors present an efficient and low cost method that allows the screening of genes or compounds activities on cell functions encompassing polarity, motility and division as well as internal compartmentation and transport. The method according to the invention lies on a precise control of focal adhesions distribution. These transmembranar complexes interact with the cytoskeleton which largely controls cell compartimentation. The accurate control of the intracellular distribution of each organelle is made possible by the use of an anisotropic adhesive pattern such as a concave adhesive pattern involving a non adhesive area. This control can also be made possible by the use of an adhesive pattern leading to a lengthening of the cell.

Problems solved by technology

These methods cannot be used when one wants to identify new genes involved in complex cell properties like protein transport, adhesion, migration, division or apoptosis, or to probe the ability of a new drug to interfere with those mechanisms.

The challenge nowadays is to associate the accuracy of modern cell biology analysis on a small number of cells to the power of high throughput automated methods on a great number of cells.

Answers to this challenge are not numerous because of several barriers: First, the cell population on the bottom of wells has a distribution which cannot be predicted.

Second, the cell shape is different from one cell to another and this parameter cannot be ignored whatever the phenotype under analysis or the quantification performed on a cell basis (intracellular localisation, number and size of particular organelles, molecular signals.).

This is particularly cumbersome.

It prevents any kind of precise analysis of the mutual distribution of intracellular compartments, or of the establishment and maintenance of cell polarity during cell division or cell migration.

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